Determinação qualitativa de enterobactérias presentes em tartarugas da Amazônia (podocnemis expansa) de vida livre e cativeiro

A tartaruga da amazônia (Podocnemis expansa) corresponde a um recurso faunístico muito importante para as populações ribeirinhas da região amazônica, além de ser uma das principais espécies indicadas para produção em cativeiro. O consumo dessa espécie como alimento na região, gerou uma demanda de estudos quanto à questão sanitária e seu impacto na saúde pública. O principal objetivo deste trabalho foi avaliar a microbiota intestinal de tartarugas da amazônia de vida livre e cativeiro, verificando a ocorrência de bactérias da Família Enterobacteriaceae no trato intestinal desses animais. Para isso, foram utilizadas 116 tartarugas adultas, de ambos os sexos, sendo que, 51 foram capturadas na Ilha de São Miguel, município de Santarém (PA), 50 animais pertenciam a um cativeiro comercial e 15 eram provenientes de um criadouro conservacionista, localizados na região metropolitana de Belém, Pará. De cada animal, foi colhida amostra de material biológico cloacal, utilizando-se swabs estéreis para em seguida serem acondicionados em tubos com meios de transporte e enviados ao laboratório para análises bacteriológicas. Todas as amostras foram imersas em caldos Selenito e BHI durante 24 horas e posteriormente semeadas em Agar Shigella-Salmonella e Agar Mac Conkey na temperatura de 37ºC por 24 horas. As UFCs (Unidades formadoras de colônia) foram semeadas em Agar Muller Hilton por mais 24 horas em estufa a 37ºC e identificadas pelo sistema Vitek® totalmente automatizado. Do total de 116 amostras foram obtidos 245 crescimentos bacterianos nos quais 83 (33,87%) eram provenientes dos animais de vida livre, com a identificação de 20 espécies bacterianas. Nos animais mantidos em cativeiro, foram obtidos 162 (65,72%) isolamentos, identificando-se 10 espécies de bactérias. Oito espécies foram encontradas em ambos os ambientes e 14 espécies em apenas um deles. A espécie Klebsiella pneumoniae foi a mais frequente, com 52 isolamentos, totalizando 21,22% dos crescimentos bacterianos, seguida de Enterobacter cloacae (35/14,29%), Serratia marcescens (29/11,84%) e Salmonella species (24/9,80%). Nos quelônios de vida livre, os microrganismos mais isolados constituiram-se dos genêros Enterobacter, Klebsiella, Citrobacter e Aeromonas. Klebsiella pneumoniae, Serratia marcescens, Enterobacter cloacae e Salmonella spp. apresentaram frequências elevadas naqueles animais cativos. Este resultado evidencia uma maior diversidade de microrganismos entre os animais de vida livre e uma contaminação elevada por amostra nos animais de cativeiro. As espécies Salmonella sp., E. coli e Acinetobacter ssp., tiveram sua frequência aumentada provavelmente devido a influência do cativeiro, sendo portanto, sugeridas como indicativas da qualidade sanitária de populações da tartaruga da Amazônia.

Bases moleculares da resistência ao mercúrio em bactérias Gram-negativas da Amazônia brasileira

Mercúrio é um dos elementos mais tóxicos tanto para seres humanos como os demais animais. Em ambientes naturais seus compostos podem ter origem em fontes naturais ou em decorrência da ação do homem. A atividade microbiana tem papel crítico na biorremediação. A resistência ao mercúrio nas bactérias está associada a um conjunto de genes organizados no operon mer. Considerando este cenário realizamos um estudo com bactérias Gram-negativas isoladas de sedimento de rios de duas áreas com distintos graus de atividade antropogênica, Barreiras e Caxiuanã. A resistência ao mercúrio foi avaliada em ensaios de crescimento in vitro da bactéria em meio contendo Hg. A presença do operon mer foi determinada por PCR para os genes RTPCABD, componentes do operon. O ensaio in vitro, determinou que apenas 2 isolados dos 107 avaliados, Acinetobacter baumannii de Caxiuanã e Pseudomonas stutzeri de Barreiras, apresentavam resistência ao Hg. Um operon mer foi identificado e caracterizado apenas do isolado Acinetobacter baumannii. Este apresenta os genes RTPCAD e sua organização e seqüência nucleotídica possui identidade total com o operon mer de um isolado de Acinetobacter calcoaceticus da Rússia.

Tropical Healthcare Epidemiology: Weather Determinants of the Etiology of Bloodstream Infections in a Brazilian Hospital

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Condições bacteriológicas da superfície das poltronas hematológicas par hemodiálise

Pós-graduação em Enfermagem (mestrado profissional) - FMB

Estudo de dinâmicas populacionais aplicado a processos de tratamento biológico de efluentes

The purpose of this work is to perform studies of mathematical modeling of the relationship of interaction occurring between microrganisms participants from wastewater treatment processes aimed at understanding, through simulations, such as inter-relationships can affect the performance of such units. The methodology was the implementation in FORTRAN computer language of mathematical models of microbial interactions. The first model addresses the interaction of bacteria-forming flakes and filamentous bacteria in activated sludge systems, which seeks to strike a balance between these bacteria to improve efficiency of the process. Another model is studied the interaction between bacteria and protozoa in activated sludge systems and analyzing the efficiency of the process, observing the changes in daily load. Microbial interactions in anaerobic reactors were dealt a third model, in which there is the mutualistic interaction between acidogenic and methanogenic bacteria. In a fourth and final model was examined the relationship between the bacteria Acinetobacter sp. and Gordonia sp., which are present in activated sludge systems, showing the competitive capacity of Acinetobacter sp. can control the growth of unwanted bacteria.

Multirresistência microbiana e opções terapêuticas disponíveis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação da atividade antimicrobiana e anti-inflamatória do extrato glicólico de Hamamelis virginiana Linnaeus

Pós-graduação em Biopatologia Bucal - ICT

Endophytic microorganisms isolated from coffee leaves, roots and branches as plant growth promoters and biocontrol agents of coffee leaf rust

Suppression of plant diseases and growth promotion due to the action of endophytic microorganisms has been demonstrated in several pathosystems. Experiments under controlled conditions involving 234 endophytic bacteria and fungi isolated from coffee leaves, roots and branches were conducted with the objective of evaluating the germination inhibition of Hemileia vastatrix urediniospores, the control of coffee leaf rust development in tests with leaf discs and on plastic bags seedling, and to promote growth of coffee seedlings. None of the fungal isolates induced plant growth or reduced disease severity. The bacterial isolates (identified by the fatty acids profile analysis) 85G (Escherichia fergusonii), 161G, 163G, 160G, 150G (Acinetobacter calcoaceticus) and 109G (Salmonella enterica) increased plant growth, the maximum being induced by 85G. This isolate produced in vitro phosphatase and indol acetic acid. In assay to control rust on coffee leaf disc, nine bacterial isolates, 64R, 137G, 3F (Brevibacillus choshinensis), 14F (Salmonella enterica), 36F (Pectobacterium carotovorum), 109G (Bacillus megaterium), 115G (Microbacterium testaceum), 116G and 119G (Cedecea davisae) significantly reduced disease severity, when applied 72 or 24h before challenging with the pathogen. In seedling tests most disease severity reduction was achieved by the isolates 109G and 119G. There was no correspondence between the organisms that promoted seedling growth and those that reduced rust severity on seedlings or leaf discs.

Etiolofia, epidemiologia e prognóstico das pneumonias associadas à ventilação mecânica em um hospital de ensino, com ênfase no estudo molecular e virulência de Staphylococcus aureus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Etiolofia, epidemiologia e prognóstico das pneumonias associadas à ventilação mecânica em um hospital de ensino, com ênfase no estudo molecular e virulência de Staphylococcus aureus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Gram-negative osteomyelitis: clinical and microbiological profile

Introduction: Despite the growing interest in the study of Gram-negative bacilli (GNB) infections, very little information on osteomyelitis caused by GNB is available in the medical literature. Objectives and methods: To assess clinical and microbiological features of 101 cases of osteomyelitis caused by GNB alone, between January 2007 and January 2009, in a reference center for the treatment of high complexity traumas in the city of Sao Paulo. Results: Most patients were men (63%), with median age of 42 years, affected by chronic osteomyelitis (43%) or acute osteomyelitis associated to open fractures (32%), the majority on the lower limbs (71%). The patients were treated with antibiotics as inpatients for 40 days (median) and for 99 days (median) in outpatient settings. After 6 months follow-up, the clinical remission rate was around 60%, relapse 19%, amputation 7%, and death 5%. Nine percent of cases were lost to follow-up. A total of 121 GNB was isolated from 101 clinical samples. The most frequently isolated pathogens were Enterobacter sp. (25%), Acinetobacter baumannii (21%) e Pseudomonas aeruginosa (20%). Susceptibility to carbapenems was about 100% for Enterobacter sp., 75% for Pseudomonas aeruginosa and 60% for Acinetobacter baumannii. Conclusion: Osteomyelitis caused by GNB remains a serious therapeutic challenge, especially when associated to nonfermenting bacteria. We emphasize the need to consider these agents in diagnosed cases of osteomyelitis, so that an ideal antimicrobial treatment can be administered since the very beginning of the therapy. (C) 2012 Elsevier Editora Ltda. All rights reserved.

Interaction of the meso-tetrakis (4-N-methylpyridyl) porphyrin with gel and liquid state phospholipid vesicles

The interaction of the cationic meso-tetrakis 4-N-methylpyridyl porphyrin (TMPyP) with large unilamellar vesicles (LUVs) was investigated in the present study. LUVs were formed by mixtures of the zwitterionic 1,2-dipalmitoyl-sn-glycero-phosphatidylcholine (DPPC) and anionic 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) phospholipids, at different DPPG molar percentages. All investigations were carried out above (50 degrees C) and below (25 degrees C) the main phase transition temperature of the LUVs (similar to 41 degrees C). The binding constant values, K-b, estimated from the time-resolved fluorescence study, showed a significant increase of the porphyrin affinity at higher mol% DPPG. This affinity is markedly increased when the LUVs are in the liquid crystalline state. For both situations, the increase of the K-b value was also followed by a higher porphyrin fraction bound to the LUVs. The displacement of the vesicle-bound porphyrins toward the aqueous medium, upon titration with the salt potassium chloride (KCl), was also studied. Altogether, our steady-state and frequency-domain fluorescence quenching data results indicate that the TMPyP is preferentially located at the LUVs Stern layer. This is supported by the zeta potential studies, where a partial neutralization of the LUVs surface charge, upon porphyrin titration, was observed. Dynamic light scattering (DLS) results showed that, for some phospholipid systems, this partial neutralization leads to the LUVs flocculation. (C) 2012 Elsevier Inc. All rights reserved.

Diversity of propanil-degrading bacteria isolated from rice rhizosphere and their potential for plant growth promotion

The herbicide propanil has long been used in rice production in southern Brazil. Bacteria isolated from contaminated soils in Massaranduba, Santa Catarina, Brazil, were found to be able to grow in the presence of propanil, using this compound as a carbon source. Thirty strains were identified as Pseudomonas (86.7%), Serratia (10.0%), and Acinetobacter (3.3%), based on phylogenetic analysis of 16S rDNA. Little genetic diversity was found within species, more than 95% homology, suggesting that there is selective pressure to metabolize propanil in the microbial community. Two strains of Pseudomonas (AF7 and AF1) were selected in bioreactor containing chemotactic growth medium, with the highest degradation activity of propanil exhibited by strain AF7, followed by AF1 (60 and 40%, respectively). These strains when encapsulated in alginate exhibited a high survival rate and were able to colonize the rice root surfaces. Inoculation with Pseudomonas strains AF7 and AF1 significantly improved the plant height of rice. Most of the Pseudomonas strains produced indoleacetic acid, soluble mineral phosphate, and fixed nitrogen. These bacterial strains could potentially be used for the bioremediation of propanil-contaminated soils and the promotion of plant growth.

Effect of low-dose gaseous ozone on pathogenic bacteria

Background: Treatment of chronically infected wounds is a challenge, and bacterial environmental contamination is a growing issue in infection control. Ozone may have a role in these situations. The objective of this study was to determine whether a low dose of gaseous ozone/oxygen mixture eliminates pathogenic bacteria cultivated in Petri dishes. Methods: A pilot study with 6 bacterial strains was made using different concentrations of ozone in an ozone-oxygen mixture to determine a minimally effective dose that completely eliminated bacterial growth. The small and apparently bactericidal gaseous dose of 20 mu g/mL ozone/oxygen (1: 99) mixture, applied for 5min under atmospheric pressure was selected. In the 2nd phase, eight bacterial strains with well characterized resistance patterns were evaluated in vitro using agar-blood in adapted Petri dishes (10(5) bacteria/dish). The cultures were divided into 3 groups: 1-ozone-oxygen gaseous mixture containing 20 mu g of O-3/mL for 5 min; 2- 100% oxygen for 5 min; 3- baseline: no gas was used. Results: The selected ozone dose was applied to the following eight strains: Escherichia coli, oxacillin-resistant Staphylococcus aureus, oxacillin-susceptible Staphylococcus aureus, vancomycin-resistant Enterococcus faecalis, extended-spectrum beta-lactamase-producing Klebsiella pneumoniae, carbapenem-resistant Acinetobacter baumannii, Acinetobacter baumannii susceptible only to carbapenems, and Pseudomonas aeruginosa susceptible to imipenem and meropenem. All isolates were completely inhibited by the ozone-oxygen mixture while growth occurred in the other 2 groups. Conclusion: A single topical application by nebulization of a low ozone dose completely inhibited the growth of all potentially pathogenic bacterial strains with known resistance to antimicrobial agents.

Expanding the Clinical and Genetic Spectrum of Human CD40L Deficiency: The Occurrence of Paracoccidioidomycosis and Other Unusual Infections in Brazilian Patients

CD40 ligand (CD40L) deficiency or X-linked hyper-IgM syndrome (X-HIGM) is a well-described primary immunodeficiency in which Pneumocystis jiroveci pneumonia is a common clinical feature. We have identified an unusual high incidence of fungal infections and other not yet described infections in a cohort of 11 X-HIGM patients from nine unrelated Brazilian families. Among these, we describe the first case of paracoccidioidomycosis (PCM) in X-HIGM. The molecular genetic analysis of CD40L was performed by gene sequencing and evaluation of CD40L protein expression. Nine of these 11 patients (82%) had fungal infections. These included fungal species common to CD40L deficiency (P. jiroveci and Candida albicans) as well as Paracoccidioides brasiliensis. One patient presented with PCM at age 11 years and is now doing well at 18 years of age. Additionally, one patient presented with a simultaneous infection with Klebsiella and Acinetobacter, and one with condyloma caused by human papilloma virus. Molecular analysis revealed four previously described CD40L mutations, two novel missense mutations (c.433 T>G and c.476 G>C) resulting in the absence of CD40L protein expression by activated CD4(+) cells and one novel insertion (c.484_485insAA) within the TNFH domain leading to a frame shift and premature stop codon. These observations demonstrated that the susceptibility to fungal infections in X-HIGM extends beyond those typically associated with X-HIGM (P. jiroveci and C. albicans) and that these patients need to be monitored for those pathogens.